Peptide–nucleotide microdroplets as a step towards a membrane-free protocell model

Koga, Shogo; Williams, David; Perriman, Adam W.; Mann, Stephen

doi:10.1038/nchem.1110

Cited by 538 publications

(714 citation statements)

References 31 publications

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“…To demonstrate that we can mimic these phase transitions, we conducted the dynamic dissolution and re‐assembly of coacervates in liposomes (Figure 2 a). As reported previously, complex coacervation can be well tuned through charge ratios,16 pH,2c temperature,17 and light 18. Herein, we show the dynamics by using complex coacervates composed of low‐complexity RNAs and short polyamines, which show reversible coacervation in response to temperature changes (higher or lower than the lower critical solution temperature (LCST)) 17.…”

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confidence: 56%

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Microfluidic Formation of Monodisperse Coacervate Organelles in Liposomes

Deng¹,

Huck²

2017

Angew Chem Int Ed

211

236

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Coacervates have been widely studied as model compartments in protocell research. Complex coacervates composed of disordered proteins and RNA have also been shown to play an important role in cellular processes. Herein, we report on a microfluidic strategy for constructing monodisperse coacervate droplets encapsulated within uniform unilamellar liposomes. These structures represent a bottom‐up approach to hierarchically structured protocells, as demonstrated by storage and release of DNA from the encapsulated coacervates as well as localized transcription.

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confidence: 56%

“…As Figure 3 b3 shows, DNA molecules partition efficiently into the artificial organelles because of electrostatic interactions and a low dielectric constant 2c, 4, 16b. DNA most likely competes with polyU for interactions with spermine in the coacervates, probably displacing some of the polyU during partitioning.…”

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confidence: 99%

Microfluidic Formation of Monodisperse Coacervate Organelles in Liposomes

Deng¹,

Huck²

2017

Angew Chem Int Ed

211

236

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“…Compartmentalization is but one characteristic, as protocells ideally also mimic the highly crowded interior of living cells, which have total macromolecule concentrations in excess of 300 g/L (7). Examples in which compartmentalization and high local concentrations are obtained concurrently, include DNA brushes (8), aqueous two-phase systems (9), and liquid coacervates (10). Phase separation or coacervation occurs in a wide range of polymer and protein solutions, often triggered by changes in temperature or salt concentration, or by the addition of coacervating agents (11).…”

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Enhanced transcription rates in membrane-free protocells formed by coacervation of cell lysate

Соколова

Spruijt

Hansen

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

305

322

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Liquid-liquid phase transitions in complex mixtures of proteins and other molecules produce crowded compartments supporting in vitro transcription and translation. We developed a method based on picoliter water-in-oil droplets to induce coacervation in Escherichia coli cell lysate and follow gene expression under crowded and noncrowded conditions. Coacervation creates an artificial cell-like environment in which the rate of mRNA production is increased significantly. Fits to the measured transcription rates show a two orders of magnitude larger binding constant between DNA and T7 RNA polymerase, and five to six times larger rate constant for transcription in crowded environments, strikingly similar to in vivo rates. The effect of crowding on interactions and kinetics of the fundamental machinery of gene expression has a direct impact on our understanding of biochemical networks in vivo. Moreover, our results show the intrinsic potential of cellular components to facilitate macromolecular organization into membranefree compartments by phase separation.microdroplets | macromolecular crowding P rotocells are minimal compartmentalized systems exhibiting key characteristics of cellular function, including metabolism and replication (1, 2). Lipid vesicles are considered the prototypical protocell as they can form functional microscopic spherical assemblies suited for in vitro gene expression (3, 4). Compartmentalization via lipid bilayers is considered essential for the emergence of cells (4), but there are alternative models based on liquid-liquid phase transitions that lead to the emergence of compartments (5, 6). Compartmentalization is but one characteristic, as protocells ideally also mimic the highly crowded interior of living cells, which have total macromolecule concentrations in excess of 300 g/L (7). Examples in which compartmentalization and high local concentrations are obtained concurrently, include DNA brushes (8), aqueous two-phase systems (9), and liquid coacervates (10). Phase separation or coacervation occurs in a wide range of polymer and protein solutions, often triggered by changes in temperature or salt concentration, or by the addition of coacervating agents (11). The (complex) coacervate droplets that are formed in such systems present macromolecularly crowded, aqueous, physical compartments, 1-100 μm in diameter (12). Recent work has identified similar liquid phase transitions in vivo in the formation of intracellular non-membrane-bound compartments exhibiting liquid-like properties, slowed down diffusion, and strongly interacting macromolecular components (13,14). Well-studied examples are the intracellular localization of DNA or RNA and proteins in Cajal bodies, P granules, and nucleoli (15-17), which can contain over 100 components. Such complexity has not been achieved in two-phase systems in vitro (18,19). Although the physics of coacervates is well understood, progress in their development as protocell models has stalled, because of the lack of demonstrations of complex biochemical proce...

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“…The resulting polymers were referred to as proteinoids, and it was shown that under certain conditions the proteinoids assembled into microscopic spheres with diameters in the micrometer range [2]. A version of microspheres was recently investigated by Koga et al [3] who created microscopic spherical compartments composed of nucleotides and a range of cationic oligopeptides or polypeptides. Unlike lipid vesicles, the microdroplets had no surface boundary structure, and their size and sedimentation properties depended on their composition.…”

Section: Introductionmentioning

confidence: 99%

Physico-chemical interactions between compartment-forming lipids and other prebiotically relevant biomolecules

Olasagasti

Maurel

Deamer

2014

BIO Web of Conferences

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Abstract. Lipids are essential constituents of contemporary living cells, serving as structural molecules that are necessary to form membranous compartments. Amphiphilic lipid-like molecules may also have contributed to prebiotic chemical evolution by promoting the synthesis, aggregation and cooperative encapsulation of other biomolecules. The resulting compartments would allow systems of molecules to be maintained that represent microscopic experiments in a natural version of combinatorial chemistry. Here we address these possibilities and describe recent results related to interactions between amphiphiles and other biomolecules during early evolution toward the first living cells.

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Peptide–nucleotide microdroplets as a step towards a membrane-free protocell model

Cited by 538 publications

References 31 publications

Microfluidic Formation of Monodisperse Coacervate Organelles in Liposomes

Microfluidic Formation of Monodisperse Coacervate Organelles in Liposomes

Enhanced transcription rates in membrane-free protocells formed by coacervation of cell lysate

Physico-chemical interactions between compartment-forming lipids and other prebiotically relevant biomolecules

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